Mab 0672

FFPE HPV58-positive (detected by PCR) cervical cancer or CIN sections (8-µm) were used for immunohistochemical staining of the HPV58-E7 protein 9 (link), anti-p16 mouse monoclonal antibody (mAb) (1:500, GT201302, Gene, Shanghai, China) and anti-Ki-67 mouse mAb (1:500, MAB-0672, MXB, Fuzhou, China). Endogenous peroxidases in the de-paraffinized sections were quenched with 0.3% hydrogen peroxide in 60% methanol for 20 min. Non-specific adsorption was minimized by incubating the sections in 2% normal goat serum (Beyotime, Shanghai, China) in PBS for 20 min. The sections were then incubated with anti-HPV58-E7 rabbit polyclonal antibody (1:500) at room temperature for 2 hours. Then, the sections were washed with PBS and incubated with an HRP-conjugated goat anti-rabbit IgG secondary or goat anti-mouse antibody (1:1000, Beyotime, Shanghai, China). The immunoreactions were visualized by incubating the sections for 3 minutes in a 0.1% 3, 30-diaminobenzidine (DAB) solution and counterstained with hematoxylin for 8 mins. The stained cells were observed under a microscope (Olympus, Tokyo, Japan).

Tissues were fixed with 4% (v/v) formaldehyde, embedded in paraffin and sectioned as described previously.37 Immunohistochemistry was performed with primary antibodies against Ki‐67 and CD34 (MAB‐0672 and Kit‐0004, MXB Biotechnologies) and an HRP–conjugated secondary antibody (GeneTech, GK500710). In negative controls, the primary antibody was replaced with 5% BSA. The stained sections were examined under a light microscope (Leica DM2000) with a digital camera (Olympus, DP73).